In 2002 a draft sequence of the mouse genome was published and the NIH described this as the genetic blueprint for the most important animal model in biomedical research. Every human gene has a mouse homologue and this enhances the probability that research using mice can provide novel insights into human disease. The long-term objective of this research program is to provide novel data concerning proteins that regulate sleep. The Specific Aims were developed to address the mission of the agency and are directly relevant to the goals of the 2007 NHLBI Strategic Research Plan. All three aims address NHLBI challenge 1.1.b. to identify intracellular targets of key signaling and transcriptional pathways in normal and pathological states. Pain disrupts sleep and there is an emerging understanding of the clinically significant interaction between sleep disruption, and pain perception. Aim 1 will test the hypothesis that B6.V-Lepob mice lacking the ability to produce the protein leptin differ from congenic C57BL/6J (B6) mice in their response to nociceptive stimuli, before and after administration of cholinergic and adenosinergic agonists into the pontine reticular formation. By replacing leptin, Aim 1 also will determine whether leptin can restore the adenosinergic modulation of nociception. Aim 2 focuses on adenosinergic and cholinergic activation of G proteins in prefrontal cortex and pontine reticular formation regions known to regulate sleep and breathing. Another family of proteins referred to as Regulators of G protein Signaling (RGS proteins) decrease the duration and strength of G protein actions. Aim 2 will use transgenic mice to test the hypothesis that cholinergic and adenosinergic activation of G proteins in sleep-related brain regions differs between mice that are homozygous and heterozygous for RGS-insensitive proteins. Aim 3 will provide functional data regarding RGS protein regulation of sleep phenotypes. Aim 3 will test the hypothesis that RGS insensitive mice exhibit genotype- specific changes in cortical acetylcholine release, EEG power, sleep, and behavior caused by cortical delivery of cholinergic and adenosinergic agonists and antagonists. The three aims are conceptually unified by their shared focus on proteins as lower-level phenotypes that modulate higher-level phenotypes of sleep, nociception, and acetylcholine release in brain regions known to regulate sleep and breathing. PUBLIC HEALTH RELEVANCE: The NIH National Center for Sleep Disorders web site notes that more than 70 million Americans have a sleep disorder. Thus, disordered sleep is a significant public health burden. This research program will provide new and needed information on brain proteins regulating sleep.